沉默 circSLAMF6 通过调控低氧微环境下胃癌细胞中 miR-204-5p/MYH9 轴抑制细胞糖酵解、迁移和侵袭。
Silencing circSLAMF6 represses cell glycolysis, migration, and invasion by regulating the miR-204-5p/MYH9 axis in gastric cancer under hypoxia.
机构信息
Department of Gastroenterology and Hepatology, Henan Provincial People's Hospital, People's Hospital of Zhengzhou University, School of Clinical Medicine, Henan University, Zhengzhou, Henan, 450003, China.
出版信息
Biosci Rep. 2020 Jun 26;40(6). doi: 10.1042/BSR20201275.
BACKGROUND
Gastric cancer (GC) is a malignant tumor of the digestive tract. Hypoxia plays an important role in the development of cancer, including GC. The present study aimed to investigate the role of circular RNA SLAMF6 (circSLAMF6) in the progression of GC under hypoxia.
METHODS
The expression of circSLAMF6, microRNA-204-5p (miR-204-5p) and myosin heavy chain 9 (MYH9) was measured by quantitative real-time polymerase chain reaction (qRT-PCR). GC cells were maintained under hypoxia (1% O2) for experiments in vitro. Glucose consumption and lactate production were determined by a Glucose Assay Kit and a Lactate Assay Kit, respectively. Levels of all protein were detected by Western blot. Cell migration and invasion were examined by Transwell assay. The interaction between miR-204-5p and circSLAMF6 or MYH9 was analyzed by dual-luciferase reporter and RNA immunoprecipitation (RIP) assays. Murine xenograft model was established to explore the role of circSLAMF6 in vivo.
RESULTS
CircSLAMF6 expression was increased in GC cells under hypoxia. Hypoxia promoted glycolysis, migration, and invasion in GC cells, which were reversed by circSLAMF6 knockdown. CircSLAMF6 was validated as a miR-204-5p sponge, and MYH9 was a target of miR-204-5p. Functionally, miR-204-5p inhibitor weakened the inhibition of circSLAMF6 knockdown on GC cell progression under hypoxia. Besides, MYH9 depletion suppressed glycolysis, migration, and invasion in GC cells under hypoxia. Importantly, circSLAMF6 deficiency inhibited tumor growth in vivo by regulating the miR-204-5p/MYH9 axis.
CONCLUSION
CircSLAMF6 was involved in glycolysis, migration, and invasion by regulating the miR-204-5p/MYH9 axis in GC cells under hypoxia.
背景
胃癌（GC）是一种消化道恶性肿瘤。缺氧在癌症的发展中起着重要作用，包括 GC。本研究旨在探讨环状 RNA SLAMF6（circSLAMF6）在缺氧下 GC 进展中的作用。
方法
通过实时定量聚合酶链反应（qRT-PCR）测量 circSLAMF6、微 RNA-204-5p（miR-204-5p）和肌球蛋白重链 9（MYH9）的表达。GC 细胞在体外维持缺氧（1% O2）进行实验。通过葡萄糖测定试剂盒和乳酸测定试剂盒分别测定葡萄糖消耗和乳酸生成。通过 Western blot 检测所有蛋白水平。通过 Transwell 测定法检测细胞迁移和侵袭。通过双荧光素酶报告和 RNA 免疫沉淀（RIP）测定分析 miR-204-5p 与 circSLAMF6 或 MYH9 的相互作用。建立小鼠异种移植模型以体内探索 circSLAMF6 的作用。
结果
缺氧下 GC 细胞中 circSLAMF6 表达增加。缺氧促进 GC 细胞的糖酵解、迁移和侵袭，circSLAMF6 敲低逆转了这些作用。circSLAMF6 被验证为 miR-204-5p 的海绵，MYH9 是 miR-204-5p 的靶标。功能上，miR-204-5p 抑制剂减弱了 circSLAMF6 敲低对缺氧下 GC 细胞进展的抑制作用。此外，缺氧下 MYH9 耗竭抑制 GC 细胞的糖酵解、迁移和侵袭。重要的是，circSLAMF6 缺乏通过调节 miR-204-5p/MYH9 轴抑制体内肿瘤生长。
结论
circSLAMF6 通过调节缺氧下 GC 细胞中的 miR-204-5p/MYH9 轴参与糖酵解、迁移和侵袭。
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